Pub Date : 2019-08-31DOI: 10.3760/CMA.J.ISSN.0254-5101.2019.08.007
Jia-fa Liu, Mi Zhang, Bihui Yang, Xuemei Deng, A. Sun, Suyun Lei
Objective To analyze the genetic structure and recombination characteristics of a newly discovered HIV-1 unique recombinant strain in Yunnan Province. Methods During a test for drug-resistant HIV genotypes in Yunnan Province in 2016, a recombinant fragment was found in the pol region of a HIV-1 strain isolated from a patient. Two overlapping segments of the HIV-1 genome were amplified by RT-PCR, and then the products were sequenced. Recombination analysis was performed using RIP, jpHMM and SimPlot3.5 software. A phylogenetic tree was constructed for homology analysis by Neighbor-joining method using MEGA6.06 software. Results A nearly full-length HIV-1 gene sequence with 8 590 bp in length was obtained. Breakpoint analysis indicated that the sequence consisted of CRF01_AE and fragments of B and C subtypes. CRF01_AE was used as the backbone with B and C subtype fragments inserted. The positions were 791 to 1 171 for CRF01_AE, 1 172 to 2 652 for C subtype fragment, 2 653 to 2 977 for B subtype fragment, and 2 978 to 9 380 for CRF01_AE using HIV-1 HXB2 as the reference strain. Conclusions Some new strains formed by cross-recombination of CRF01_AE and B and C subtypes were discovered in Yunnan Province in recent years. It was found that the recombination pattern of the newly discovered strain was complex, suggesting that close attention should be paid to the changes in epidemic trends, which was of great importance to understand the current prevalence and epidemic trends of HIV-1. Key words: Human immunodeficiency virus type 1; Unique recombinant form; Near-full-length gene sequence analysis
{"title":"Analysis of the nearly full-length genome of a novel HIV-1 CRF01_AE/B/C recombinant strain isolated in Yunnan Province","authors":"Jia-fa Liu, Mi Zhang, Bihui Yang, Xuemei Deng, A. Sun, Suyun Lei","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.08.007","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.08.007","url":null,"abstract":"Objective \u0000To analyze the genetic structure and recombination characteristics of a newly discovered HIV-1 unique recombinant strain in Yunnan Province. \u0000 \u0000 \u0000Methods \u0000During a test for drug-resistant HIV genotypes in Yunnan Province in 2016, a recombinant fragment was found in the pol region of a HIV-1 strain isolated from a patient. Two overlapping segments of the HIV-1 genome were amplified by RT-PCR, and then the products were sequenced. Recombination analysis was performed using RIP, jpHMM and SimPlot3.5 software. A phylogenetic tree was constructed for homology analysis by Neighbor-joining method using MEGA6.06 software. \u0000 \u0000 \u0000Results \u0000A nearly full-length HIV-1 gene sequence with 8 590 bp in length was obtained. Breakpoint analysis indicated that the sequence consisted of CRF01_AE and fragments of B and C subtypes. CRF01_AE was used as the backbone with B and C subtype fragments inserted. The positions were 791 to 1 171 for CRF01_AE, 1 172 to 2 652 for C subtype fragment, 2 653 to 2 977 for B subtype fragment, and 2 978 to 9 380 for CRF01_AE using HIV-1 HXB2 as the reference strain. \u0000 \u0000 \u0000Conclusions \u0000Some new strains formed by cross-recombination of CRF01_AE and B and C subtypes were discovered in Yunnan Province in recent years. It was found that the recombination pattern of the newly discovered strain was complex, suggesting that close attention should be paid to the changes in epidemic trends, which was of great importance to understand the current prevalence and epidemic trends of HIV-1. \u0000 \u0000 \u0000Key words: \u0000Human immunodeficiency virus type 1; Unique recombinant form; Near-full-length gene sequence analysis","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"601-607"},"PeriodicalIF":0.0,"publicationDate":"2019-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48889102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-08-31DOI: 10.3760/CMA.J.ISSN.0254-5101.2019.08.011
Yanan Hou
Norovirus is one of the leading causes of acute non-bacterial gastroenteritis. Emergence of new genotypes due to frequent genetic recombination and antigenic drift increases the prevalence of norovirus infection worldwide. Norovirus infection has become a global public health concern. This review focused on disease burden, vaccine research, cell culture and animal models regarding norovirus infection. Key words: Norovirus; Gastroenteritis; Vaccine
{"title":"Progress in norovirus research and vaccine development","authors":"Yanan Hou","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.08.011","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.08.011","url":null,"abstract":"Norovirus is one of the leading causes of acute non-bacterial gastroenteritis. Emergence of new genotypes due to frequent genetic recombination and antigenic drift increases the prevalence of norovirus infection worldwide. Norovirus infection has become a global public health concern. This review focused on disease burden, vaccine research, cell culture and animal models regarding norovirus infection. \u0000 \u0000 \u0000Key words: \u0000Norovirus; Gastroenteritis; Vaccine","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"628-632"},"PeriodicalIF":0.0,"publicationDate":"2019-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41873367","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-08-31DOI: 10.3760/CMA.J.ISSN.0254-5101.2019.08.002
Xiaodong Liu, Haoqiu Wang, X. Lou, W. Zheng, Zhibei Zheng, Tao Liu, Wei Zhang, Qi Chen, Shi Cheng
Objective To investigate the genomic characteristics and virulence factors of emetic-type Bacillus cereus strains isolated from food in Hangzhou for better understanding their pathogenic potential. Methods Real-time PCR was performed to detect the ces gene cluster (cereulide) in 132 Bacillus cereus strains isolated from food from 2015 to 2017. Genomes of cereulide-positive strains were sequenced using Illumina MiSeq sequencing platform. Genome annotation, virulence factor detection, comparative and evolutionary analysis were performed after the sequences of genomes were assembled. Results Twelve strains (9.09%) carried the ces gene. Their genome sizes ranged from 5.35 to 5.75 Mb and GC contents from 35.25 to 35.43 mol%. All of them harbored the full cereulide biosynthesis gene cluster, nonhemolytic enterotoxin (NHE)-encoding gene cluster (nheA, nheB and nheC) and hemolysin Ⅲ (hlyⅢ). The average nucleotide identity (ANI) between the 12 isolates and the reference strain NC7401 (Accession number: AP007209) was over 99.35%. Phylogenetic analysis demonstrated these strains were clustered into the same branch with local clinical isolates and the emetic-type Bacillus cereus strains of NC7401 and AH187. Conclusions The genomic sequences of the emetic-type Bacillus cereus strains isolated from food in Hangzhou area were highly similar to that of the reference strain NC7401. Results of the genomic analysis suggested that these isolates carried many virulence factors that were related to pathogenicity. Key words: Bacillus cereus; Genome sequencing; Virulence factor; Genomic analysis
{"title":"Analysis of genomic characteristics and virulence factors of emetic-type Bacillus cereus strains isolated in Hangzhou","authors":"Xiaodong Liu, Haoqiu Wang, X. Lou, W. Zheng, Zhibei Zheng, Tao Liu, Wei Zhang, Qi Chen, Shi Cheng","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.08.002","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.08.002","url":null,"abstract":"Objective \u0000To investigate the genomic characteristics and virulence factors of emetic-type Bacillus cereus strains isolated from food in Hangzhou for better understanding their pathogenic potential. \u0000 \u0000 \u0000Methods \u0000Real-time PCR was performed to detect the ces gene cluster (cereulide) in 132 Bacillus cereus strains isolated from food from 2015 to 2017. Genomes of cereulide-positive strains were sequenced using Illumina MiSeq sequencing platform. Genome annotation, virulence factor detection, comparative and evolutionary analysis were performed after the sequences of genomes were assembled. \u0000 \u0000 \u0000Results \u0000Twelve strains (9.09%) carried the ces gene. Their genome sizes ranged from 5.35 to 5.75 Mb and GC contents from 35.25 to 35.43 mol%. All of them harbored the full cereulide biosynthesis gene cluster, nonhemolytic enterotoxin (NHE)-encoding gene cluster (nheA, nheB and nheC) and hemolysin Ⅲ (hlyⅢ). The average nucleotide identity (ANI) between the 12 isolates and the reference strain NC7401 (Accession number: AP007209) was over 99.35%. Phylogenetic analysis demonstrated these strains were clustered into the same branch with local clinical isolates and the emetic-type Bacillus cereus strains of NC7401 and AH187. \u0000 \u0000 \u0000Conclusions \u0000The genomic sequences of the emetic-type Bacillus cereus strains isolated from food in Hangzhou area were highly similar to that of the reference strain NC7401. Results of the genomic analysis suggested that these isolates carried many virulence factors that were related to pathogenicity. \u0000 \u0000 \u0000Key words: \u0000Bacillus cereus; Genome sequencing; Virulence factor; Genomic analysis","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"572-577"},"PeriodicalIF":0.0,"publicationDate":"2019-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45366183","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-08-31DOI: 10.3760/CMA.J.ISSN.0254-5101.2019.08.001
Jiangqing Huang, Zhi-chang Zhao, Yao Chen, Yingping Cao, Bin Li
Objective To analyze the molecular characteristics of qnrS-positive Escherichia coli (E.coli) strains resistant to quinolone. Methods A total of 57 qnrS1-positive clinical isolates were collected from Fujian Medical University Union Hospital. Plasmid-mediated quinolone resistance (PMQR) genes [qnrA, qnrB, qnrC, qnrD, aac(6′)-Ib-cr, qepA and oqxAB] and β-lactamase genes (blaCTX-M-1, blaCTX-M-2, blaCTX-M-8, blaCTX-M-9, blaSHV and blaTEM) were detected by PCR and then sequenced. Agar dilution method was used to analyze the antimicrobial susceptibility of the qnrS1-positive strains. Phylogenetic analysis was conducted using PCR. Multilocus sequence typing (MLST) was performed for phenotyping. Enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) was used to evaluate the genetic similarity between those isolates. Transferability of the qnrS1 genes carried by the 57 strains was examined by conjugation test with the sodiumazide-resistant E. coli J53 as the recipient strain. Mutations in the quinolone resistance-determining regions (QRDR) in those strains were analyzed by PCR. Results All of the qnrS1-positive E. coli strains showed high resistance to quinolones. PMQR genes were harbored by 14 (24.6%) isolates. Extended spectrum β-lactamases (ESBLs)-producing isolates accounted for 68.4%. Mutations in the QRDR of gyrA, gyrB, parC and parE genes were found in 56 (98.2%) strains and the most frequent point mutations were S83L (89.5%) in gyrA gene, S80I (54.4%) in parC gene and P415V (28.1%) in parE gene. The qnrS1 gene was successful transferred from 13 (22.8%) isolates to E. coli J53 by conjugation. Five plasmid incompatibility groups were detected. Phylogenetic analysis showed that there were 36 (63.2%), 13 (22.8%), 1 (1.8%) and 7 (12.3%) isolates belonging to groups A, B1, B2 and D, respectively. The 57 qnrS1-positive E. coli strains were assigned to 50 ERIC types and 39 sequence types (ST) based on the results of ERIC-PCR and MLST. Conclusions Mutations in the QRDR in E. coli strains were associated with qnrS1 gene and might play a critical role in the dissemination of quinolone-resistant bacteria. Key words: qnrS; Escherichia coli; Quinolone resistance; Plasmid-mediated quinolone resistance; Quinolone resistance-determining region
{"title":"Molecular characteristics of qnrS1-positive Escherichia coli resistant to quinolone","authors":"Jiangqing Huang, Zhi-chang Zhao, Yao Chen, Yingping Cao, Bin Li","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.08.001","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.08.001","url":null,"abstract":"Objective \u0000To analyze the molecular characteristics of qnrS-positive Escherichia coli (E.coli) strains resistant to quinolone. \u0000 \u0000 \u0000Methods \u0000A total of 57 qnrS1-positive clinical isolates were collected from Fujian Medical University Union Hospital. Plasmid-mediated quinolone resistance (PMQR) genes [qnrA, qnrB, qnrC, qnrD, aac(6′)-Ib-cr, qepA and oqxAB] and β-lactamase genes (blaCTX-M-1, blaCTX-M-2, blaCTX-M-8, blaCTX-M-9, blaSHV and blaTEM) were detected by PCR and then sequenced. Agar dilution method was used to analyze the antimicrobial susceptibility of the qnrS1-positive strains. Phylogenetic analysis was conducted using PCR. Multilocus sequence typing (MLST) was performed for phenotyping. Enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) was used to evaluate the genetic similarity between those isolates. Transferability of the qnrS1 genes carried by the 57 strains was examined by conjugation test with the sodiumazide-resistant E. coli J53 as the recipient strain. Mutations in the quinolone resistance-determining regions (QRDR) in those strains were analyzed by PCR. \u0000 \u0000 \u0000Results \u0000All of the qnrS1-positive E. coli strains showed high resistance to quinolones. PMQR genes were harbored by 14 (24.6%) isolates. Extended spectrum β-lactamases (ESBLs)-producing isolates accounted for 68.4%. Mutations in the QRDR of gyrA, gyrB, parC and parE genes were found in 56 (98.2%) strains and the most frequent point mutations were S83L (89.5%) in gyrA gene, S80I (54.4%) in parC gene and P415V (28.1%) in parE gene. The qnrS1 gene was successful transferred from 13 (22.8%) isolates to E. coli J53 by conjugation. Five plasmid incompatibility groups were detected. Phylogenetic analysis showed that there were 36 (63.2%), 13 (22.8%), 1 (1.8%) and 7 (12.3%) isolates belonging to groups A, B1, B2 and D, respectively. The 57 qnrS1-positive E. coli strains were assigned to 50 ERIC types and 39 sequence types (ST) based on the results of ERIC-PCR and MLST. \u0000 \u0000 \u0000Conclusions \u0000Mutations in the QRDR in E. coli strains were associated with qnrS1 gene and might play a critical role in the dissemination of quinolone-resistant bacteria. \u0000 \u0000 \u0000Key words: \u0000qnrS; Escherichia coli; Quinolone resistance; Plasmid-mediated quinolone resistance; Quinolone resistance-determining region","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"565-571"},"PeriodicalIF":0.0,"publicationDate":"2019-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46313601","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-08-31DOI: 10.3760/CMA.J.ISSN.0254-5101.2019.08.003
Xiaoxia Li, H. Lu, Zhiqun Zhang, G. Zhuo
Objective To investigate the effects of calcitonin gene-related peptide (CGRP) on the inflammatory response induced by lipopolysaccharide (LPS) of Klebsiella pneumoniae. Methods Klebsiella pneumoniae was cultured in vitro to extracted LPS. Different concentrations of LPS were used to stimulate BEAS-2B cells. The activation of human β defensin 2 (hBD-2) in these cells was detected by immunofluorescence assay before and after adding different concentrations of CGRP. MTT assay and flow cytometry were respectively used to detect the proliferation and apoptosis of BEAS-2B cells after LPS stimulation with and without CGRP treatment. Neutrophil granules released in the cells after CGRP treatment were detected using neutrophil gelatinase-related apolipoprotein (NGAL) as the marker. Results Immunofluorescence assay results showed that LPS at different concentrations could significantly increase the relative expression of hBD-2 in BEAS-2B cells, which was significantly inhibit by CGRP intervention. LPS at 50 ng/ml, 100 ng/ml and 200 ng/ml had no significant effect on the activity of BEAS-2B cells, while treatment with 400 ng/ml of LPS for 24 h could significantly reduce the activity and promote the apoptosis of BEAS-2B cells. In addition, remarkedly increased cell activity and suppressed cell apoptosis were induced when BEAS-2B cells were treated with 10 nmol/L of CGRP in combination with LPS. LPS at different concentrations could induce the release of neutrophil-specific granules, while the LPS-induce release could be significantly inhibited by 10 nmol/L of CGRP. Conclusions CGRP could inhibit the expression of hBD-2, promote cell proliferation and reduce the degranulation of neutrophils to down-regulate the inflammatory response induced by LPS of Klebsiella pneumoniae. Key words: Klebsiella pneumoniae; Calcitonin gene-related peptide; βdefensin; Inflammatory response
{"title":"Calcitonin gene-related peptide down-regulates the inflammatory response induced by lipopolysaccharide of Klebsiella pneumoniae","authors":"Xiaoxia Li, H. Lu, Zhiqun Zhang, G. Zhuo","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.08.003","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.08.003","url":null,"abstract":"Objective \u0000To investigate the effects of calcitonin gene-related peptide (CGRP) on the inflammatory response induced by lipopolysaccharide (LPS) of Klebsiella pneumoniae. \u0000 \u0000 \u0000Methods \u0000Klebsiella pneumoniae was cultured in vitro to extracted LPS. Different concentrations of LPS were used to stimulate BEAS-2B cells. The activation of human β defensin 2 (hBD-2) in these cells was detected by immunofluorescence assay before and after adding different concentrations of CGRP. MTT assay and flow cytometry were respectively used to detect the proliferation and apoptosis of BEAS-2B cells after LPS stimulation with and without CGRP treatment. Neutrophil granules released in the cells after CGRP treatment were detected using neutrophil gelatinase-related apolipoprotein (NGAL) as the marker. \u0000 \u0000 \u0000Results \u0000Immunofluorescence assay results showed that LPS at different concentrations could significantly increase the relative expression of hBD-2 in BEAS-2B cells, which was significantly inhibit by CGRP intervention. LPS at 50 ng/ml, 100 ng/ml and 200 ng/ml had no significant effect on the activity of BEAS-2B cells, while treatment with 400 ng/ml of LPS for 24 h could significantly reduce the activity and promote the apoptosis of BEAS-2B cells. In addition, remarkedly increased cell activity and suppressed cell apoptosis were induced when BEAS-2B cells were treated with 10 nmol/L of CGRP in combination with LPS. LPS at different concentrations could induce the release of neutrophil-specific granules, while the LPS-induce release could be significantly inhibited by 10 nmol/L of CGRP. \u0000 \u0000 \u0000Conclusions \u0000CGRP could inhibit the expression of hBD-2, promote cell proliferation and reduce the degranulation of neutrophils to down-regulate the inflammatory response induced by LPS of Klebsiella pneumoniae. \u0000 \u0000 \u0000Key words: \u0000Klebsiella pneumoniae; Calcitonin gene-related peptide; βdefensin; Inflammatory response","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"578-582"},"PeriodicalIF":0.0,"publicationDate":"2019-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44390024","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-08-31DOI: 10.3760/CMA.J.ISSN.0254-5101.2019.08.005
Wei-Zheng Song, Xiangyu Chen, Y. Shao, Y. Hao, Ying Wang
Objective To express HIV-1 capsid p24 antigen in an eukaryotic expression system and to evaluate its antigenicity and potential in the early diagnosis of HIV. Methods The full-length gene of HIV-1 p24 and the signal peptide DRVI gene were amplified by PCR. The signal peptide DRVI preceding the p24 gene was introduced using fusion PCR, and cloned into vector pDRVI1.0. Two recombinant plasmids pDRVI-p24 and pDRVI-p24s were constructed and transfected into 293F cells. Expression and secretion of p24 protein were detected by SDS-PAGE, Ni-NTA column chromatography and molecular sieve were used to purify p24s protein. The purified protein was identified by Western blot and indirect ELISA using human/mouse HIV-1-positive serum samples. Results The eukaryotic expression system for HIV-1 p24 antigen was successfully established with high efficiency. The target protein of interest with the signal peptide DRVI was obviously detected in the supernatants of cell culture. The recombinant protein had good specificity and sensitivity based on the results of serological tests using serum samples of five HIV-1-positive and five HIV-negative mice, 30 HIV-1-positive patients and 50 HIV-1-negative healthy individuals. Conclusions The eukaryotic expression system for HIV-1 p24 antigen was successfully established. The purified HIV-1 p24s antigen had good antigenicity. An indirect ELISA assay with good specificity and sensitivity for the detection of HIV-1 was preliminarily constructed and showed great potential for application. Key words: HIV-1; p24 core antigen; Eukaryotic expression; Serological diagnosis
{"title":"Eukaryotic expression and serodiagnostic potential of HIV-1 p24 antigen","authors":"Wei-Zheng Song, Xiangyu Chen, Y. Shao, Y. Hao, Ying Wang","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.08.005","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.08.005","url":null,"abstract":"Objective \u0000To express HIV-1 capsid p24 antigen in an eukaryotic expression system and to evaluate its antigenicity and potential in the early diagnosis of HIV. \u0000 \u0000 \u0000Methods \u0000The full-length gene of HIV-1 p24 and the signal peptide DRVI gene were amplified by PCR. The signal peptide DRVI preceding the p24 gene was introduced using fusion PCR, and cloned into vector pDRVI1.0. Two recombinant plasmids pDRVI-p24 and pDRVI-p24s were constructed and transfected into 293F cells. Expression and secretion of p24 protein were detected by SDS-PAGE, Ni-NTA column chromatography and molecular sieve were used to purify p24s protein. The purified protein was identified by Western blot and indirect ELISA using human/mouse HIV-1-positive serum samples. \u0000 \u0000 \u0000Results \u0000The eukaryotic expression system for HIV-1 p24 antigen was successfully established with high efficiency. The target protein of interest with the signal peptide DRVI was obviously detected in the supernatants of cell culture. The recombinant protein had good specificity and sensitivity based on the results of serological tests using serum samples of five HIV-1-positive and five HIV-negative mice, 30 HIV-1-positive patients and 50 HIV-1-negative healthy individuals. \u0000 \u0000 \u0000Conclusions \u0000The eukaryotic expression system for HIV-1 p24 antigen was successfully established. The purified HIV-1 p24s antigen had good antigenicity. An indirect ELISA assay with good specificity and sensitivity for the detection of HIV-1 was preliminarily constructed and showed great potential for application. \u0000 \u0000 \u0000Key words: \u0000HIV-1; p24 core antigen; Eukaryotic expression; Serological diagnosis","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"591-595"},"PeriodicalIF":0.0,"publicationDate":"2019-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42781981","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-08-31DOI: 10.3760/CMA.J.ISSN.0254-5101.2019.08.008
Linlin Zhu, Zhixuan Xu, Mingming Zhang
Objective To investigate the effects of glaucocalyxin A (GLA) on the expression of signal transduction and activator of transcription 3 (STAT3) signaling pathway in HCCLM3 cells for better understanding the role of GLA in tumor metastasis. Methods HCCLM3 cells were stimulated by different concentrations (0.1 ng/ml, 1 ng/ml, 10 ng/ml, 100 ng/ml and 1 000 ng/ml) of GLA for 48 h. Expression of IL-6, STAT3/pSTAT3Tyr705 and MMP9 at mRNA and protein levels were detected by qRT-PCR and Western blot, respectively. Transwell invasion and migration assays were performed to analyze cell invasion and migration. ELISA was used to measure the concentrations of IL-6 in culture media. Results Expression of IL-6 at mRNA and protein levels, STAT3Tyr705 phosphorylation, and the migration and invasion of HCCLM3 cells were significantly enhanced after stimulation of HCCLM3 cells with 0.1 ng/ml of GLA, but inhibited by GLA at the concentration of 1 000 ng/ml. Moreover, more IL-6 was secreted out of the HCCLM3 cells treated with 0.1 ng/ml of GLA. Conclusions GLA might affect the metastasis of HCCLM3 cells through regulating the expression of IL-6 and the phosphorylation of STAT3Tyr705. These regulatory effects were closely related to the concentration of GLA. Key words: Glaucocalyxin A; Interleukin-6; Signal transduction and activator of transcription 3; Hepatocarcinoma
{"title":"Glaucocalyxin A regulates STAT3 signaling pathway through influencing IL-6 expression in HCCLM3 cells","authors":"Linlin Zhu, Zhixuan Xu, Mingming Zhang","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.08.008","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.08.008","url":null,"abstract":"Objective \u0000To investigate the effects of glaucocalyxin A (GLA) on the expression of signal transduction and activator of transcription 3 (STAT3) signaling pathway in HCCLM3 cells for better understanding the role of GLA in tumor metastasis. \u0000 \u0000 \u0000Methods \u0000HCCLM3 cells were stimulated by different concentrations (0.1 ng/ml, 1 ng/ml, 10 ng/ml, 100 ng/ml and 1 000 ng/ml) of GLA for 48 h. Expression of IL-6, STAT3/pSTAT3Tyr705 and MMP9 at mRNA and protein levels were detected by qRT-PCR and Western blot, respectively. Transwell invasion and migration assays were performed to analyze cell invasion and migration. ELISA was used to measure the concentrations of IL-6 in culture media. \u0000 \u0000 \u0000Results \u0000Expression of IL-6 at mRNA and protein levels, STAT3Tyr705 phosphorylation, and the migration and invasion of HCCLM3 cells were significantly enhanced after stimulation of HCCLM3 cells with 0.1 ng/ml of GLA, but inhibited by GLA at the concentration of 1 000 ng/ml. Moreover, more IL-6 was secreted out of the HCCLM3 cells treated with 0.1 ng/ml of GLA. \u0000 \u0000 \u0000Conclusions \u0000GLA might affect the metastasis of HCCLM3 cells through regulating the expression of IL-6 and the phosphorylation of STAT3Tyr705. These regulatory effects were closely related to the concentration of GLA. \u0000 \u0000 \u0000Key words: \u0000Glaucocalyxin A; Interleukin-6; Signal transduction and activator of transcription 3; Hepatocarcinoma","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"608-612"},"PeriodicalIF":0.0,"publicationDate":"2019-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44286390","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-08-31DOI: 10.3760/CMA.J.ISSN.0254-5101.2019.08.012
Xi Yang, X. Bai, Yan-mei Xu
Shiga toxin (Stx), which can be divided into Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2), is an important virulence factor of Shigella spp. and certain strains of Escherichia coli. Stx, encoded by λ-like phage, blocks protein synthesis through removal of an adenine residue from the 28S rRNA. Stx can also induce apoptosis through multiple pathways. Humans may suffer from diarrhea, hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS) and even death when infected with Shiga toxin-producing bacteria. At present, there is no specific treatment for diseases caused by Stx. In recent years, the application of Stx in cancer therapy and imaging has aroused great interest. This review provided a brief overview of Stx in its nomenclature, typing, structure, genetics, pathogenesis and application perspectives. Key words: Shiga toxin; Shigella dysenteriae; Shiga toxin-producing Escherichia coli; λ-like phage
{"title":"Advance in Shiga toxin","authors":"Xi Yang, X. Bai, Yan-mei Xu","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.08.012","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.08.012","url":null,"abstract":"Shiga toxin (Stx), which can be divided into Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2), is an important virulence factor of Shigella spp. and certain strains of Escherichia coli. Stx, encoded by λ-like phage, blocks protein synthesis through removal of an adenine residue from the 28S rRNA. Stx can also induce apoptosis through multiple pathways. Humans may suffer from diarrhea, hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS) and even death when infected with Shiga toxin-producing bacteria. At present, there is no specific treatment for diseases caused by Stx. In recent years, the application of Stx in cancer therapy and imaging has aroused great interest. This review provided a brief overview of Stx in its nomenclature, typing, structure, genetics, pathogenesis and application perspectives. \u0000 \u0000 \u0000Key words: \u0000Shiga toxin; Shigella dysenteriae; Shiga toxin-producing Escherichia coli; λ-like phage","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"633-637"},"PeriodicalIF":0.0,"publicationDate":"2019-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47231926","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-08-31DOI: 10.3760/CMA.J.ISSN.0254-5101.2019.08.006
Guangming Zhang, Jianyi Mai, Zejun Wang, Sheng-li Meng, Z. Pang, Meng′an Chen, S. Shen
Objective To detect and analyze enteroviruses causing suspected aseptic meningitis in a kindergarten in Jinhua City, Zhejiang Province. Methods Viral RNA was extracted from samples and cDNA was prepared by reverse transcription. PCR was performed to amplify the partial sequences of the 5′-untranslated region (UTR) and VP1 gene of enteroviruses. Serotypes of the viruses were determined by comparing the homology between the partial sequences of VP1 gene. Phylogenetic tree of the partial VP1 sequences was constructed using MEGA6. Results This study included seven patients and twenty-six asymptomatic students. Coxsakievirus A10 (CV-A10) was detected in 48.5% of the students and echovirus 6 (Echo 6) in 21.2%. Besides, 12.1% of the students might be co-infected by the two viruses. Among the seven patients, six were infected by CV-A10 and the other one might have co-infection. According to the phylogenetic analysis, CV-A10 strains detected in this study were closely related to those isolated in China in recent years, including the strains isolated in Xiamen in 2015 and Yunnan in 2017, while the Echo 6 strains were phylogenetically related to those isolated in Yunnan, Guangzhou and Shandong in 2014. Conclusions CV-A10 and Echo 6 were detected in the cases with suspected aseptic meningitis and had close phylogenetic relationships to the strains appeared in China in recent years. Key words: Aseptic meningitis; Enterovirus; Coxsackievirus A10; Echovirus 6
{"title":"Molecular detection and sequence analysis of enteroviruses isolated from a suspected outbreak of aseptic meningitis","authors":"Guangming Zhang, Jianyi Mai, Zejun Wang, Sheng-li Meng, Z. Pang, Meng′an Chen, S. Shen","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.08.006","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.08.006","url":null,"abstract":"Objective \u0000To detect and analyze enteroviruses causing suspected aseptic meningitis in a kindergarten in Jinhua City, Zhejiang Province. \u0000 \u0000 \u0000Methods \u0000Viral RNA was extracted from samples and cDNA was prepared by reverse transcription. PCR was performed to amplify the partial sequences of the 5′-untranslated region (UTR) and VP1 gene of enteroviruses. Serotypes of the viruses were determined by comparing the homology between the partial sequences of VP1 gene. Phylogenetic tree of the partial VP1 sequences was constructed using MEGA6. \u0000 \u0000 \u0000Results \u0000This study included seven patients and twenty-six asymptomatic students. Coxsakievirus A10 (CV-A10) was detected in 48.5% of the students and echovirus 6 (Echo 6) in 21.2%. Besides, 12.1% of the students might be co-infected by the two viruses. Among the seven patients, six were infected by CV-A10 and the other one might have co-infection. According to the phylogenetic analysis, CV-A10 strains detected in this study were closely related to those isolated in China in recent years, including the strains isolated in Xiamen in 2015 and Yunnan in 2017, while the Echo 6 strains were phylogenetically related to those isolated in Yunnan, Guangzhou and Shandong in 2014. \u0000 \u0000 \u0000Conclusions \u0000CV-A10 and Echo 6 were detected in the cases with suspected aseptic meningitis and had close phylogenetic relationships to the strains appeared in China in recent years. \u0000 \u0000 \u0000Key words: \u0000Aseptic meningitis; Enterovirus; Coxsackievirus A10; Echovirus 6","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"596-600"},"PeriodicalIF":0.0,"publicationDate":"2019-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41661272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-08-31DOI: 10.3760/CMA.J.ISSN.0254-5101.2019.08.013
Qiao Li, Yuepeng Wang, Yusen Zhou, Z. Kou
Toll-like receptor 4 (TLR4), a type Ⅰ transmembrane protein, has been extensively studied in the Toll-like receptor family at present. TLR4 ligands include lipopolysaccharides (LPS) present in the outer membrane of gram-negative bacteria and monophosphoryl lipid A (MPLA) which is a derivative of LPS. TLR4 agonists, alone, as a major component of compound adjuvants or in combination with other TLRs agonists, have been widely used as adjuvants in various vaccines and demonstrated great potential in vaccine development. This review addressed the discovery, application, features and prospect of novel vaccine adjuvants based on TLR4 agonists, aiming to provide reference for rational use of adjuvants and further development. Key words: Adjuvant; Immunostimulant; TLR4; Vaccine
{"title":"Advances in novel vaccine adjuvants based on TLR4 agonists","authors":"Qiao Li, Yuepeng Wang, Yusen Zhou, Z. Kou","doi":"10.3760/CMA.J.ISSN.0254-5101.2019.08.013","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.0254-5101.2019.08.013","url":null,"abstract":"Toll-like receptor 4 (TLR4), a type Ⅰ transmembrane protein, has been extensively studied in the Toll-like receptor family at present. TLR4 ligands include lipopolysaccharides (LPS) present in the outer membrane of gram-negative bacteria and monophosphoryl lipid A (MPLA) which is a derivative of LPS. TLR4 agonists, alone, as a major component of compound adjuvants or in combination with other TLRs agonists, have been widely used as adjuvants in various vaccines and demonstrated great potential in vaccine development. This review addressed the discovery, application, features and prospect of novel vaccine adjuvants based on TLR4 agonists, aiming to provide reference for rational use of adjuvants and further development. \u0000 \u0000 \u0000Key words: \u0000Adjuvant; Immunostimulant; TLR4; Vaccine","PeriodicalId":10089,"journal":{"name":"Chinese journal of microbiology and immunology","volume":"39 1","pages":"638-644"},"PeriodicalIF":0.0,"publicationDate":"2019-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44789817","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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